Clearly mark your weeks. Could have better captions and more thorough analysis. -UM Week 5&6
10/20/2013
A master plate of 8 of the colonies was made and placed into the incubator for overnight incubation.
I will go in tomorrow morning and spin down the samples in the shake incubator and midi prep.
Picture of the plate will be uploaded tomorrow morning as well as a picture of the 8 samples
10/20/2013
Image of the sample A plate with clones that grew using the cleaned up PCR products and cleaned up pNIC.
There were ~12 colonies that crew on the plate (LB,Kanamycin, Sucrose). Sample A plate is shown above. A 2:4 concentration (vector:insert) was used while preparing the plate. These colonies will be used to make a master plate to allow the colonies to grow even more.
10/19/13
The first round of cloning resulted in orangish/white colonies that were distorted and smeared across the plate. These plates are thought to have been made improperly because this happened to others who used the same plates as well. Plate A and Plate B cloning was unsuccessful. I will make new plates today and check the new plates in the morning.
-Inserts and vectors were made to clone using the PCR cleaned up PNIC and PCR squared products.
Nano drop of pNIC Bsa-4 cut
The concentration of the cut pNIC was decent. I used it to go ahead and clone my DNA
Nano drop of STPP T.Cruzi PCR Squared products
The concentration of the cleaned up PCR Squared products was usable. I am going to go ahead and use it for the first round of cloning.
Week 3 & 4
Grant - you need more updates here (RE digest, Tail primer design, etc.) . Also - this image doesn't seem correct - Dr. B 092513
PCR Squared Successful:
Secondary PCR Successful:
Secondary PCRs (failures):
RE digest:
Primary PCR:
Picture of gel electrophoresis obtained from the primary PCR. Lane 1 is empty. Lane 2 contains 100 bp DNA ladder. Lane 3 is empty. Lane 4 contains primary PCR solution with oligo mix, dNTP, 5X rxn buffer, Q5 Polymerase dH20, and blue juice stain.
Week 1 & 2:
Grant - Submit to DNA, Oligo Primer result Dr. B 090913
NanoDrop-
PCR pGBR22-
PCR gel of pGBR22 purple protein
Analyse DNA:
Determining the DNA sequence of the pGBR22 plasmid.
Week 5&6
10/20/2013
A master plate of 8 of the colonies was made and placed into the incubator for overnight incubation.
- I will go in tomorrow morning and spin down the samples in the shake incubator and midi prep.
Picture of the plate will be uploaded tomorrow morning as well as a picture of the 8 samples10/20/2013
Image of the sample A plate with clones that grew using the cleaned up PCR products and cleaned up pNIC.
There were ~12 colonies that crew on the plate (LB,Kanamycin, Sucrose). Sample A plate is shown above. A 2:4 concentration (vector:insert) was used while preparing the plate. These colonies will be used to make a master plate to allow the colonies to grow even more.
10/19/13
The first round of cloning resulted in orangish/white colonies that were distorted and smeared across the plate. These plates are thought to have been made improperly because this happened to others who used the same plates as well. Plate A and Plate B cloning was unsuccessful. I will make new plates today and check the new plates in the morning.
-Inserts and vectors were made to clone using the PCR cleaned up PNIC and PCR squared products.
Nano drop of pNIC Bsa-4 cut
The concentration of the cut pNIC was decent. I used it to go ahead and clone my DNA
Nano drop of STPP T.Cruzi PCR Squared products
The concentration of the cleaned up PCR Squared products was usable. I am going to go ahead and use it for the first round of cloning.
Week 3 & 4
Grant - you need more updates here (RE digest, Tail primer design, etc.) . Also - this image doesn't seem correct - Dr. B 092513
PCR Squared Successful:
Secondary PCR Successful:
Secondary PCRs (failures):
RE digest:
Primary PCR:
Week 1 & 2:
Grant - Submit to DNA, Oligo Primer result Dr. B 090913
NanoDrop-
PCR pGBR22-
Analyse DNA:
Determining the DNA sequence of the pGBR22 plasmid.
Virtual Gels-
Submitting DNA to DNA Sequencing Facility: